| Summary: | 碩士 === 國立中興大學 === 獸醫學系 === 82 === The reverse transcription-polymerase chain reaction ( RT-PCR )
was used to detect Infectious bursal disease virus ( IBDV )
RNA. In the part of IBDV genome,two sets of primes detect
standard strain. The specific primers were decided to the
presented nucleotide sequence analysis of genome segment A of
IBDV with artificial synthesis. The cDNA was synthesized at
$45^\circ C$ ,60 minutes. The first amplification was proceeded
30 cycles with $92^\circ C 1$ minute, $35^\circ C$ 1 minute,
$72^\circ C 2$ minutes. The second amplification condition is
the same as the first amplification condition. The RT-PCR
product was observed on 2.5\% agarose contained ethidium
bromide . RT-PCR product were identified by using restriction
endonucleases.The two sets of primers were selected and used as
the detection of IBDV RNA. The sensitivity test showed that the
lower limit of detection was approximately $10^{1.5} TCID_{50
}$ of IBDV . The primers did not react with nucleic acids of
chicken bursal cell, chicken spleen cell, chicken embryo
fibroblast cell, fowlpox virus, Marek's disease virus, turkey
herpes virus, avain reovirus, avain influenza virus, infectious
bronchitis and infectious laryngotracheitis virus. In addition
to yhe detection of IBDV RNA of bursas, buffycoat collected
from the chickens experimentally exposed to IBDV, the primers
were also used to detect the viral RNA extracted from spleens
of spadger and chicken bursas in several poultry farms where
the IBD infections were suspected.
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